Single-cell RNA sequencing analysis reveals the relationship of bone marrow and osteopenia in STZ-induced type 1 diabetic mice

Jinjie Zhong; Xingjia Mao; Heyangzi Li; Gerong Shen; Xi Cao; Ning He; Jingyu Wang; Lintao Xu; Jun Chen; Xinghui Song; Shuangshuang Liu; Xiaoming Zhang; Yueliang Shen; Lin-Lin Wang; Chuan Xiang; Ying-Ying Chen

doi:10.1016/j.jare.2022.01.006

Single-cell RNA sequencing analysis reveals the relationship of bone marrow and osteopenia in STZ-induced type 1 diabetic mice

J Adv Res. 2022 Nov:41:145-158. doi: 10.1016/j.jare.2022.01.006. Epub 2022 Jan 21.

Authors

Jinjie Zhong¹, Xingjia Mao², Heyangzi Li³, Gerong Shen⁴, Xi Cao³, Ning He⁴, Jingyu Wang⁵, Lintao Xu⁵, Jun Chen¹, Xinghui Song⁶, Shuangshuang Liu⁶, Xiaoming Zhang³, Yueliang Shen⁷, Lin-Lin Wang⁸, Chuan Xiang⁹, Ying-Ying Chen¹⁰

Affiliations

¹ Department of Basic Medicine Sciences, and Department of Obstetrics of the Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310058, China.
² Department of Orthopedic, The Second Hospital of Shanxi Medical University, Taiyuan 030001, China.
³ Department of Basic Medicine Sciences, and Department of Emergency Medicine, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou 310058, China.
⁴ Department of Basic Medicine Sciences, and Department of Orthopaedics of Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou 310058, China.
⁵ Department of Neurosurgery, Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou 310009, China.
⁶ Core Facilities, Zhejiang University School of Medicine, Hangzhou 310058, China.
⁷ Department of Basic Medicine Sciences, Zhejiang University School of Medicine, Hangzhou 310058, China.
⁸ Department of Basic Medicine Sciences, and Department of Orthopaedics of Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou 310058, China. Electronic address: wanglinlin@zju.edu.cn.
⁹ Department of Orthopedic, The Second Hospital of Shanxi Medical University, Taiyuan 030001, China. Electronic address: chuanxiang@sxmu.edu.cn.
¹⁰ Department of Basic Medicine Sciences, and Department of Obstetrics of the Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310058, China. Electronic address: bchenyy@zju.edu.cn.

Abstract

Introduction: Type 1 diabetes (T1D) is a multifactorial autoimmune disease. Broad knowledge about the genetics, epidemiology and clinical management of T1D has been achieved, but understandings about the cell varieties in the bone marrow during T1D remain limited.

Objectives: We aimed to present a profile of the bone marrow cells and reveal the relationship of bone marrow and osteopenia in streptozotocin (STZ)-induced T1D mice.

Methods: The whole bone marrow cells from the femurs and tibias of healthy (group C) and STZ-induced T1D mice (group D) were collected for single-cell RNA sequencing analysis. Single-cell flow cytometry and immunohistochemistry were performed to confirm the proportional changes among bone marrow neutrophils (BM-neutrophils) (Cxcr2⁺, Ly6g⁺) and B lymphocytes (Cd19⁺). X-ray and micro-CT were performed to detect bone mineral density. The correlation between the ratio of BM-neutrophils/B lymphocytes and osteopenia in STZ-induced T1D mice was analyzed by nonparametric Spearman correlation analysis.

Results: The bone marrow cells in groups C and D were divided into 12 clusters, and 249 differentially expressed genes were found. The diversity of CD45⁺ immune cells between groups C and D were greatly affected: the proportion of BM-neutrophils showed a significant increase while the proportion of B lymphocytes in group D showed a significant decrease. X-ray and micro-CT analyses confirmed that osteopenia occurred in group D mice. In addition, the results of single-cell flow cytometry and correlation analysis showed that the ratio of BM-neutrophils/B lymphocytes negatively correlated with osteopenia in STZ-induced T1D mice.

Conclusion: A single-cell RNA sequencing analysis revealed the profile and heterogeneity of bone marrow immune cells in STZ-induced T1D mice for the first time. The ratio of BM-neutrophils/B lymphocytes negatively correlated with osteopenia in STZ-induced T1D mice, which may enhance understanding for treating T1D and preventing T1D-induced osteopenia.

Keywords: Bone marrow; Lymphocyte; Neutrophil; Osteopenia; Single-cell RNA sequencing; Type 1 diabetes.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Bone Diseases, Metabolic*
Bone Marrow
Diabetes Mellitus, Experimental*
Diabetes Mellitus, Type 1*
Mice
Sequence Analysis, RNA
Streptozocin

Substances

Streptozocin